Abstract
Adenovirus is a widely used vector for cancer gene therapy because of its high infection efficiency and capacity for transgene expression in both dividing and nondividing cells. However, neutralisation of adenovirus by pre-existing antibodies can lead to inefficient delivery, and the wide tissue distribution of the coxsackie and adenovirus receptor (CAR, the primary receptor for adenovirus type 5) precludes target selectivity. These limitations have largely restricted therapeutic use of adenovirus to local or direct administration. A successful viral gene therapy vector would be protected from neutralising antibodies and exhibit a preferential tropism for target cells. We report here the development of a covalent coating and retargeting strategy using a multivalent hydrophilic polymer based on poly-[N-(2-hydroxypropyl)metha- crylamide] (pHPMA). Incorporation of targeting ligands such as basic fibroblast growth factor and vascular endothelial growth factor on to the polymer-coated virus produces ligand-mediated, CAR-independent binding and uptake into cells bearing appropriate receptors. Retargeted virus is resistant to antibody neutralisation and can infect receptor-positive target cells selectively in mixed culture, and also in xenografts in vivo. Multivalent polymeric modification of adenovirus is an effective way of changing its tropism and interaction with the immune system. As a non-genetic one-step process, the technology is simple, versatile and should yield vectors with an improved safety profile.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Bergelson JM et al. Isolation of a common receptor for coxsackie B viruses and adenoviruses 2 and 5 Science 1997 275: 1320–1323
Chirmule N et al. Immune responses to adenovirus and adeno-associated virus in humans Gene Therapy 1999 6: 1574–1583
Stallwood Y, Fisher KD, Gallimore PH, Mautner V . Neutralisation of adenovirus infectivity by ascitic fluid from ovarian cancer patients Gene Therapy 2000 7: 637–643
Yang Y, Li Q, Ertl HCJ, Wilson JM . Cellular and humoral immune responses to viral antigens create barriers to lung-directed gene therapy with recombinant adenoviruses J Virol 1995 69: 2004–2015
Searle PF, Mautner V . Adenoviral vectors: not to be sneezed at Gene Therapy 1998 5: 725–727
Henry LJ et al. Characterization of the knob domain of the adenovirus type 5 fiber protein expressed in Escherichia coli J Virol 1994 68: 5239–5246
Michael SI, Hong JS, Curiel DT, Engler JA . Addition of a short peptide ligand to the adenovirus fiber protein Gene Therapy 1995 2: 660–668
Reynolds PN, Dimitriev I, Curiel DT . Insertion of an RGD motif into the HI loop of adenovirus fiber protein alters the distribution of transgene expression of the systemically administered vector Gene Therapy 1999 6: 1336–1339
Wickham TJ et al. Targeted adenovirus gene-transfer to endothelial and smooth-muscle cells by using bispecific antibodies J Virol 1996 70: 6831–6838
Miller CR et al. Differential susceptibility of primary and established human glioma cells to adenovirus infection: targeting via the epidermal growth factor receptor achieves fiber receptor-independent gene transfer Cancer Res 1998 58: 5738–5748
Gu DL et al. Fibroblast growth factor 2 retargeted adenovirus has redirected cellular tropism: evidence for reduced toxicity and enhanced antitumor activity in mice Cancer Res 1999 59: 2608–2614
Rihova B . Receptor-mediated targeted drug or toxin delivery Adv Drug Del Rev 1998 29: 273–289
Vasey PA et al. Phase I clinical and pharmacokinetic study of PK1 [N-(2- hydroxypropyl)methacrylamide copolymer doxorubicin]: first member of a new class of chemotherapeutic agents – drug-polymer conjugates Clin Cancer Res 1999 5: 83–94
Dash PR et al. Decreased binding to proteins and cells of polymeric gene delivery vectors surface-modified with a multivalent hydrophilic polymer J Biol Chem 2000 275: 3793–3802
Fisher KD et al. A versatile system for receptor-mediated gene delivery permits increased entry of DNA into target cells, enhanced delivery to the nucleus and elevated rates of transgene expression Gene Therapy 2000 7: 1337–1343
O'Riordan CR et al. PEGylation of adenovirus with retention of infectivity and protection from neutralizing antibody in vitro and in vivo Hum Gene Ther 1999 10: 1349–1358
Romanczuk H et al. Modification of an adenoviral vector with biologically selected peptides: a novel strategy for gene delivery to cells of choice Hum Gene Ther 1999 10: 2615–2626
Wickham TJ, Mathias P, Cheresh DA, Nemerow GR . Integrins αvβ3 and αvβ5 promote adenovirus internalization but not virus attachment Cell 1993 73: 309–319
Greber UF, Willetts M, Webster P, Helenius A . Stepwise dismantling of adenovirus 2 during entry into cells Cell 1993 75: 477–486
Cotten M, Weber J . The adenovirus protease is required for virus entry into host cells Virology 1995 213: 494–502
Greber UF et al. The role of the nuclear pore complex in adenovirus DNA entry EMBO J 1997 16: 5998–6007
Fredman JN, Engler JA . Adenovirus precursor to terminal protein interacts with the nuclear matrix in vivo and in vitro J Virol 1993 67: 3384–3395
Doukas J et al. Retargeted delivery of adenoviral vectors through fibroblast growth factor receptors involves unique cellular pathways Faseb J 1999 13: 1459–1466
Bikfalvi A et al. Interaction of vasculotropin vascular endothelial-cell growth-factor with human umbilical vein endothelial-cells–binding, internalization, degradation and biological effects J Cell Physiol 1991 149: 50–59
Mayr GA, Freimuth P . A single locus on human chromosome 21 directs the expression of a receptor for adenovirus type 2 in mouse A9 cells J Virol 1997 71: 412–418
Croyle MA, Yu QC, Wilson JM . Development of a rapid method for the PEGylation of adenovirus with enhanced transduction and improved stability under harsh storage conditions Hum Gene Ther 2000 11: 1713–1722
Chillon M, Lee JH, Fasbender A, Welsh MJ . Adenovirus complexed with polyethylene glycol and cationic lipid is shielded from neutralizing antibodies in vitro Gene Therapy 1998 5: 995–1002
Lemay P, Boudin M, Mileville M, Boulanger P . Human adenovirus type 2 protein IIIA. Purification and characterisation Virology 1980 101: 131–143
Rihova B et al. Biocompatibility of N-(2-hydroxypropyl) methacrylamide copolymers containing adriamycin – immunogenicity, and effect on hematopoietic stem cells in bone marrow in vivo and mouse splenocytes and human peripheral blood lymphocytes in vitro Biomaterials 1989 10: 335–342
Stocks SJ, Jones AJM, Ramey CW, Brooks DE . A fluorometric assay of the degree of modification of protein primary amines with polyethylene-glycol Anal Biochem 1986 154: 232–234
Acknowledgements
We are grateful to Paul Freimuth for supplying A9 and A9–21 cells, to David Oupicky for synthesis of 14C-pHPMA-ONp, to Ellis Muggleton and Helen Ketteringham for producing the Ad5 knob domain protein, to Kim Ward for expert assistance with flow cytometry and to Alan Rickinson, Phillip Gallimore and Lawrence Young for critical reading of the manuscript. AdgfpΔE1 and Adβ-galΔE1 were constructed by VM in collaboration with Peter Searle. This work was supported by the Cancer Research Campaign (LWS and YS), the Medical Research Council (VM and KDF) and AstraZeneca plc (KDF).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Fisher, K., Stallwood, Y., Green, N. et al. Polymer-coated adenovirus permits efficient retargeting and evades neutralising antibodies. Gene Ther 8, 341–348 (2001). https://doi.org/10.1038/sj.gt.3301389
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.gt.3301389
Keywords
This article is cited by
-
Challenges and progress toward tumor-targeted therapy by systemic delivery of polymer-complexed oncolytic adenoviruses
Cancer Gene Therapy (2022)
-
Molecular retargeting of antibodies converts immune defense against oncolytic viruses into cancer immunotherapy
Nature Communications (2019)
-
White paper on microbial anti-cancer therapy and prevention
Journal for ImmunoTherapy of Cancer (2018)
-
Barriers to systemic application of virus-based vectors in gene therapy: lessons from adenovirus type 5
Virus Genes (2017)
-
Aptamer-facilitated Protection of Oncolytic Virus from Neutralizing Antibodies
Molecular Therapy - Nucleic Acids (2014)